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Control of cellular redox levels

An oxidative and bacterial technology, applied in the direction of extracellular fluid diseases, medical raw materials derived from bacteria, antibacterial drugs, etc., can solve problems such as sepsis, excessive cell activation response, etc.

Pending Publication Date: 2018-08-21
伊丽莎白麦克纳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Aberrant TLR signaling responses may lead to exaggerated cellular activation responses leading to sepsis

Method used

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  • Control of cellular redox levels
  • Control of cellular redox levels
  • Control of cellular redox levels

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0183] Example 1: Preparation of active ingredients for TLR agonist compositions

[0184] Examples of how to formulate the compositions include the following procedures.

[0185] active ingredient. The active ingredients are derived from bacterial fermentation and cell isolation processes as described below. Lactobacillus delbrueckii, ssp. bulgaricus (referred to herein as L. bulgaricus) is the organism used in this example, but the bacterial organism can be any Gram-positive or Gram-positive Negative bacteria or both.

[0186] Lactobacillus delbrueckii ssp bulgaricus contains 1.5% casein hydrolyzate, 1% yeast extract, 3% lactose, 0.2% sodium acetate, 0.02% sodium formate, 0.01% disodium 5-inosinate , 0.01% manganese sulphate, 0.05% magnesium sulphate and 0.05% polysorbate 80, pH 6.4 growth medium for fermentation. The inoculated medium was incubated at 37°C until the fermentation reached steady growth, as determined by metabolic arrest. The broth was cooled to 4°C and ha...

Embodiment 2

[0190] Example 2: TLR Screening Assays

[0191] Toll-like receptor (TLR) stimulation was tested by assessing NF-κB activation in HEK293 cells expressing a given TLR or Nod-like receptor (NLR). In seven different human TLRs (TLR2, 3, 4, 5, 7, 8 and 9 (Invivogen, San Diego, CA)) and in two different human NLRs (TLRs containing a nucleotide-binding oligomerization domain The activity of the samples was tested on proteins 1 and 2 (NOD1 and NOD2)). Each ligand was tested on TLR or NLR cells at a final concentration of 1 / 100 stock solution and compared to a control ligand as described below. This step was repeated three times.

[0192] The control ligands, control cell lines and samples used in the examples are shown in Table 1.

[0193] Table 1: Control Ligand and Control Cell Line Information for Ligand Screening Tests

[0194]

[0195]

[0196] usual procedure. TLR stimulation in the screen was tested by assessing NF-κB activation in HEK293 cells expressing a given TLR...

Embodiment 3

[0198] Example 3: Effect of Process Variables on Changes in TLR Signaling

[0199] By observing the effect of certain process changes on TLR signaling, it was noted that TLR signaling patterns may be altered.

[0200] image 3 Differences in cell morphology with TLR signaling of Gram-positive organisms are shown. Lysates from Pediococcus acidilactici (a coccus organism) produced higher TLR4 and lower NOD2 signals compared to the bacillus organism L. bulgaricus, which (P acidilactici) was only slightly higher than the TLR2 signal produced by L. bulgaricus. NOD2 activation was significantly reduced in P. acidilactici, reflecting the lower muramyl peptides found in cocci.

[0201] Such as Figure 4 It was shown that TLR signaling patterns are also affected by the time of culture harvest of Bacillus coagulans lysates. Bacillus coagulans was incubated at 45° C. and 250 rpm using shake flasks containing standard yeast extract / glucose medium with a pH value of 6.5. The harvest ...

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Abstract

Disclosed herein are compositions and methods for regulating redox status and / or reducing oxidative stress in a subject, the methods and compositions comprising TLR agonists comprising bacterial lysates and / or lysate fractions. Also disclosed are compositions and methods comprising bacterial lysates and / or lysate fractions formulated or administered in combination with one or more other therapeutic or pharmaceutical agents.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Application No. 62 / 253,542, filed November 10, 2015. This application is also a continuation-in-part of U.S. Application No. 14 / 640,075, filed March 6, 2015, which is a continuation-in-part of U.S. Application No. 14 / 034,044, filed September 23, 2013, which claimed the September 21, 2012 Priority to U.S. Provisional Application No. 61 / 704,090 filed on . This application is also a continuation-in-part of US Application No. 13 / 743,194, filed January 16, 2013, which claims priority to US Provisional Application No. 61 / 586,975, filed January 16, 2012. All of the aforementioned applications are hereby incorporated by reference in their entirety. Background technique [0003] The innate immune response is one of the ways to regulate inflammation. Inflammation is stimulated by chemical factors released by damaged cells and serves to create a physical barrier against the spre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/74A61P39/00
CPCA61K35/74A61P1/00A61P1/02A61P1/04A61P1/08A61P1/12A61P1/14A61P1/16A61P1/18A61P11/00A61P11/06A61P13/10A61P13/12A61P15/00A61P15/10A61P17/00A61P17/02A61P17/06A61P19/02A61P19/10A61P21/00A61P25/00A61P25/04A61P25/14A61P25/16A61P25/20A61P25/24A61P25/26A61P25/28A61P27/02A61P27/12A61P27/16A61P29/00A61P3/00A61P3/04A61P31/04A61P31/18A61P35/00A61P35/02A61P3/06A61P37/02A61P37/06A61P37/08A61P39/00A61P39/06A61P43/00A61P7/00A61P7/02A61P9/00A61P9/04A61P9/06A61P9/10A61P9/12A61P3/10Y02A50/30
Inventor 伊丽莎白·麦克纳
Owner 伊丽莎白麦克纳
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